Age-dependent physiological responses of last instarHelicoverpa zea larvae to oral and topical application of precocene II

Newly-ecdysed last instar larvae ofH. zea grouped into 100-, 200-, 300-, or 400-mg categories were fed diet containing precocene II or given precocene II topically on the abdomen. The time for larvae to reach a maximal weight, time to pupation, growth rate, and the amount of precocene II excreted were calculated. Younger larvae of lower weights, which were fed or topically treated with precocene II required more time to reach their maximal weight, had a lower maximal weight, a lower growth rate, and required more time to pupate than control larvae. Older larvae represented by the largest weight category were less sensitive to precocene II, had a shorter delay in reaching maximal weight, and a shorter delay in the time to pupation than control larvae; larvae in the largest weight category that were fed precocene II also had smaller decreases in the growth rate. Growth rate declines for larvae given topical doses of precocene II, however, were largest for the oldest larvae. All larvae given a single topical dose excreted precocene II for several days and were most efficient at eliminating smaller doses; larger, older larvae excreted more precocene II than smaller, younger larvae. Age-dependent responses to precocene II indicate that growth and metabolic processes, as well as xenobiotic metabolism, change in last instar larvae.     

Behavioral changes and growth inhibition in last instar larvae of Heliothis zea induced by oral and topical application of precocene II

In diet choice tests, newly-ecdysed fifth instar Heliothis zea larvae randomly chose diet containing 4 mM precocene II or diet without precocene II but initially preferred to feed on diet without precocene II. After two days, however, they were found more frequently on diet containing precocene II. Nevertheless, larval growth and development were inhibited when forced to eat diet containing precocene II. Removal of the maxillae, the site of the primary gustatory receptors, did not prevent the growth inhibition caused by larval consumption of diet with precocene II. Topical treatments with precocene II also caused larval growth inhibition, and daily treatments were more detrimental to the larvae than a large single dose. Addition of methylenedioxyprecocene (6,7-methylenedioxy-2,2-dimethylchromene) to the diet did not reverse the observed larval growth inhibition and this compound was itself significantly toxic. These results indicate that larval growth and development are disrupted after precocene II ingestion, but not because of preingestive discrimination by olfaction and gustation.     

Utilization of Δ5,7 - and Δ8-sterols by larvae of Heliothis zea

Larvae from two populations of Heliothis zea were reared on artificial diets containing various sterols, which supported suboptimal growth, and their tissue sterols were characterized in order to determine how these dietary sterols are utilized by this insect. The sterols studied included Δ^5,7-sterols (7-dehydrocholesterol or ergosterol), Δ⁸-sterols (lanosterol and/or 24-dihydrolanosterol), and a Δ⁵-sterol (4,4-dimethylcholesterol). Although larvae did not develop on 4,4-dimethylcholesterol, those fed primarily Δ⁸-4,4,14-trimethylsterols developed to the third instar. When the latter sterols were spared with cholesterol, the larvae reached the sixth instar and contained 4,4,14-trimethylsterols as well as cholesterol in their tissues. When larvae were fed 7-dehydrocholesterol, <1% of the larvae from one population developed to the sixth instar and these larvae contained 7-dehydrocholesterol as their principal sterol. The other larvae successfully completed their larval stage when they were transferred from the diet containing 7-dehydrocholesterol (or no sterol) to a diet containing cholesterol within at least 9 days. The sterol composition of larvae transferred from a diet containing cholesterol to a diet containing 7-dehydrocholesterol, after they had reached 60% of their final weight, was 54% cholesterol and 46% 7-dehydrocholesterol. The major sterol isolated from the tissues of the larvae fed ergosterol was also 7-dehydrocholesterol. Therefore, although the larva of H. zea can dealkylate and saturate the side chain of the Δ^5,7,22-24β-methylsterol, it carries out little metabolism of the B ring of the nucleus. These studies demonstrate that, when Δ^5,7- or Δ⁸-sterols are the principal sterols in the diet of H. zea, they are absorbed and incorporated into its tissues, although they slow the rate of growth and may prevent complete development of the larva.     

Toxicity and disruption of midgut physiology in larvae of the European corn borer,Ostrinia nubilalis,by anion transporter blockers

In this study, four blockers of anion transporters (ATs) belonging to four different classes of organic acids, including DIDS (4, 4'‐diisothiocyanatostilbene‐2, 2'‐ disulfonic acid; a stilbene disulfonic acid), NPPB [(5‐nitro‐2‐(3‐phenylpropylamino) benzoic acid; an anthranilic acid)], 9‐AC (anthracene‐9‐carboxylic acid; an aromatic carboxylic acid), and IAA‐94 (indanyloxy acetic acid; an indanyloxy alkanoic acid), were tested for their toxicity against the European corn borer (ECB), Ostrinia nubilalis. All the AT blockers inhibited the growth of larvae, increased the developmental time, and decreased survival compared to controls, when second‐instar ECB larvae were fed for seven days on treated diet. In general, DIDS and NPPB were the most active compounds, with the rank order of activity being DIDS>NPPB>IAA‐94>9‐AC. All the AT blockers decreased the midgut alkalinity in fifth‐instar larvae when fed for 3 h on treated diet. Effective concentrations required for 50% decrease in midgut alkalinity (EC₅₀) ranged between 29.1 and 41.2 ppm and the rank order of activity was NPPB>DIDS>IAA‐94>9‐AC. Similarly, all the tested AT blockers inhibited ³⁶Cl^? uptake from the midgut lumen in fifth‐instar larvae when fed for 3 h on treated diet. Concentrations required for 50% inhibition of ³⁶Cl^? uptake (IC₅₀) ranged between 7.4 and 11.0 ppm and the rank order of activity was DIDS>NPPB>9‐AC >IAA‐94. Modest to highly strong positive correlations observed among growth, midgut alkalinity, and midgut Cl^? ion transport in AT blocker–fed larvae suggested that these effects are causally related to each other. Finally, AT blockers have the potential to become good candidates for development of insecticides with a unique mode of action. © 2009 Wiley Periodicals, Inc.     

Sequestration of host plant carotenoids in the larval tissues of Helicoverpa zea

To determine the cause of the unique yellow coloration in mandibular glands of soybean-fed Helicoverpa zea larvae, the accumulation of carotenoids in various tissues of last instar larvae fed soybean, cotton and tomato foliage was quantified. Five carotenoids were detected in the foliage of all host plants but at significantly different concentrations. Xanthophylls rather than carotenes were most likely to accumulate in larval tissues. Carotenoids accumulated at different rates and some were significantly affected by larval diet. Highest levels of carotenoid accumulation, notably lutein, were detected in the testes, followed by midgut epithelium, fat body and integument. The midgut epithelium contained the greatest and the testes the least diversity of carotenoid types. Low levels of lutein were detected in both labial and mandibular glands. Tomato foliage had the highest carotenoid content and caterpillar tissues fed these leaves often had the highest amounts of carotenoid. However, the accumulation of carotenoids did not protect larvae from antibiotic effects of tomato foliage because these caterpillars had the highest mortality and slowest growth rates of all the three host plants. Transport and absorption of lipid and oxidative stress may be some reasons for differential carotenoid accumulation.     

The effect of GNA lectin on the α-amylase activity of the beet armyworm,Spodoptera exigua Hb. (Lepidoptera: Noctuidae)

Beet armyworm (Spodoptera exigua Hb.) (Lepidoptera: Noctuidae) is the major pest of sugar beet (Beta vulgaris). Pesticide applications are the main method of the insect control. So, alternative method/s is/are needed to control this insect species. So, in the current study, the effect of Galanthus nivalis agglutinin (GNA) (snowdrop lectin) on beet armyworm α-amylase was studied. Measurement of the amylase activity of the larval midgut fed on artificial diet and sugar beet leaves showed that the enzyme activity was higher when the larvae fed on artificial diet. However, in both cases, the fourth instar larvae had the greatest amylase activity. Thus, fourth instar larvae were offered artificial diet containing 1 and 2% GNA. Both treatments of the lectin significantly reduced the α-amylase activity of the insect. For example, amylase activity of the fourth instar larvae in the control (fed only on artificial diet) was 2.62 Uml^?1 whilst the activity of the enzyme in the two treatments including diet containing 2 and 1% lectin was 1.45 and 1.75 Uml^?1, respectively. The achieved data showed that lectin, in addition to have toxic effect on the larval growth and development, affects the α-amylase activity of the insect gut.     

EFFECTS OF BT RICE ON THE FOOD CONSUMPTION,GROWTH AND SURVIVAL OF CHILO SUPPRESSAUS LARVAE UNDER DIFFERENT TEMPERATURES*

Abstract The Bt transgenic rice line, i.e. KMD1 containing a synthetic cry 1 Ab gene from Bacillus thuringiensis Berliner was tested in the laboratory for evaluating its effects on larval food consumption, growth and survival of striped stem borer (SSB), Chilo suppressalis (Walker) under different temperatures. The food consumption and body weight growth as well as the survival of both 3rd and 5th instar larvae fed on Bt rice were significantly reduced, as compared with those on the control. The temperature showed no marked effect on both food consumption and body weight growth of larvae fed on Bt rice stems since the onset of 3rd instar, conversely had significant effects as since the initial stage of 5th instar. In contrast, the temperature did not distinctly influence the survival of both 3rd and 5th instar larvae. There was a linearly positive relationship between the corrected mortality of larvae and the accumulative amount of Bt rice tissues ingested by larvae.     

Endogenous and induced monooxygenase activity in gypsy moth larvae feeding on natural and artificial diets

NADPH oxidase activity was measured in third to sixth instar gypsy moth larvae fed oak or pine foliage. Activity levels ranged from 400 to 1,900 pmol NADPH oxidized/min/mg microsomal protein, but enzyme activity was not correlated with host plant ingested. Similarly, activity levels in larvae fed diets containing inducers, such as the terpenoid α-pinene or pentamethylbenzene, ranged from 700 to 1,500 pmol NADPH oxidized/min/mg protein, levels that were comparable to those measured for larvae fed control diets. O-demethylase activity in older instar gypsy moth larvae fed pine averaged 109 pmol p-nitrophenol/min/mg protein, and activity levels in those fed diet containing α-pinene ranged from 22 to 55 pmol/min/mg protein. Although statistically significant, these induced O-demethylase levels are well below those observed for Heliothis zea larvae. Our findings indicate that monooxygenases play a minor, if any, role in the ability of later instar gypsy moth larvae to develop successfully on pine foliage.     

Interaction of Acronicta rumicis (Lepidoptera: Noctuidae) and its larval parasitoid, Glyptapanteles liparidis (Hymenoptera: Ichneumonidae)

As a result of parasitism by Glyptapanteles liparidis in the first, second, third and fourth instar larvae of Acronicta rumicis, the mortality of each larval stage was found to be 46.67, 90, 71 and 16.67%, respectively. The mortality was highest when G. liparidis parasitized the second and third instar larvae. The difference in mortality between the parasitized group and the control group was 72.14% in the second instar larvae. With regards to the food consumption of the parasitized larvae, the first and second instar larvae consumed 6495.58 ± 646.52 mm² (leaf surface) and 7951.12 ± 4167.36 mm², respectively, while the third and fourth larvae consumed 13 826.77 ± 3396.66 mm² and 18 599.85 mm², respectively, showing that food consumption increased with instar stages of the host larvae. The clutch size of G. liparidis increased in relation to the instar stages of the host: it was 25.25 ± 7.89, 48.65 ± 53.75, 91.09 ± 44.52 and 114 individuals when they were fed with the first, second, third and the fourth instar larvae of the host, respectively.     

Responses of representative midgut detoxifying enzymes from Heliothis zea and Spodoptera frugiperda to trichothecenes

The relative levels of representative midgut detoxifying enzymes were measured in last instar larvae of Heliothis zea and Spodoptera frugiperda after the insects had been fed for 48 h on diets containing representative trichothecenes, which are protein synthesis inhibitors. 4-Nitroanisole O-demethylation was induced (as indicated by increased enzyme activity) by 1.6-fold in H. zea and by 6.1-fold in S. frugiperda when fed 250 ppm of deoxynivalenol, a dose that severely retarded growth of the insects. Induction of this activity was also noted with lower levels of deoxynivalenol in both species, and with diacetoxyscirpenol and T-2 toxin in H. zea. 1-Chloro-2,4-dinitrobenzene glutathione transferase conjugation was induced by c. 1.3-fold with deoxynivalenol at 250 ppm and 25 ppm, and with T-2 toxin at 25 ppm in H. zea; but was slightly inhibited (c. 10–20%) in S. frugiperda in some cases.1-Naphthyl acetate hydrolysis was generally unaffected by all trichothecenes tested at all doses in H. zea, but was inhibited by c. 30% in S. frugiperda by lower doses of all three trichothecenes. However, a new 1-naphthyl acetate esterase band was noted after exposure to 250 ppm of deoxynivalenol, and to some extent lower doses of deoxynivalenol and the other trichothecenes in both insect species. The hydrolysis of a radiolabeled model trichothecene, 4-monoacetoxyscripenol, was induced by 3.1-fold in H. zea and 2.7-fold in S. frugiperda with 250 ppm of deoxynivalenol (the only compound and dose tested). This activity could be inhibited by nearly 100% with 10⁻⁴ M paraoxon, suggesting a serine-hydroxyl esterase was involed. Although this activity could not be recovered from gels, indirect competitive inhibition assays with 1-naphthyl acetate suggested the induced band could be responsible for some of the hydrolysis of the 4-monoacetoxyscirpenol.     

Entomopathogenic fungi disturbed the larval growth and feeding performance of Ocinara varians (Lepidoptera: Bombycidae) larvae

Abstract Feeding experiments using three strains of entomopathogenic fungi, Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea were conducted with newly moulted 3rd–5th instar Ocinara varians Walker larvae in the laboratory. The mortality of larvae immersed individually in spore suspension (1 × 10⁷ spores/mL) of all the strains was ≥ 80% except 5th instar larvae treated with M. anisopliae which transformed into pupae, but did not result in adult emergence. The growth (total body mass), consumption, relative consumption rate and relative growth rate, were reduced at all three larval stages, while developmental time was extended in infected larvae with concurrent significant increase in approximate digestibility in infected larvae. Conversion of digested food (ECD) and ingested food (ECI) values declined in infected larvae as compared to the healthy larvae (control). The 5th instar larvae treated with M. anisopliae showed higher ECD and ECI values than control. Based on mortality and growth inhibition it can be suggested that all the studied fungal strains have a high potential for biocontrol and could be developed into biocontrol agents against O. varians.     

取食不同食料植物对安婀珍蝶的营养利用及中肠四种酶活力的影响

分别用薇甘菊Mikania micrantha HBK、飞机草Chromolaena odorata L.、革命菜Gynura crepidioides Benth、菜心Brassica parachinensis Bailey和白菜Brassica chinensis L.饲养安婀珍蝶Actinote anteas,计算各龄期试虫的存活率;测定不同食料植物对安婀珍蝶4龄幼虫的营养利用以及中肠中淀粉酶、海藻糖酶、蔗糖酶和羧酸酯酶活性的影响。结果表明：薇甘菊饲养的试虫,其4、5、6龄的存活率高于其他供试食料植物上的试虫;取食薇甘菊的安婀珍蝶4龄幼虫相对生长率和食物利用率明显高于取食其他3种食料植物的试虫,但其相对取食量明显较低;取食薇甘菊的安婀珍蝶4龄试虫羧酸酯酶比活力明显高于以其他3种植物为食的试虫;取食白菜的4龄试虫,其蔗糖酶、海藻糖酶和淀粉酶比活力较低。这些研究结果表明安婀珍蝶对革命菜、菜心和白菜存在不同程度的取食利用障碍,由此判定他们不会成为安婀珍蝶的寄主植物。     

Co‐expression of the proteinase inhibitors oryzacystatin I and oryzacystatin II in transgenic potato alters Colorado potato beetle larval development

Colorado potato beetle (CPB; Leptinotarsa decemlineata Say, Coleoptera: Chrysomelidae) has shown a remarkable adaptability to a variety of control measures. Although oryzacystatin I and II (OCI and OCII) have potential in controlling pests that use cysteine proteinases for food digestion, expression of a single OC gene in potato exhibited a minimal or no effect on CPB fitness traits. The aim of this study was to examine the effect of coexpressed OCI and OCII in potato (Solanum tuberosum L.) cultivars Desiree, Draga?evka and Jelica on CPB larvae. Growth parameters, consumption rates and food utilization, as well as activity of proteases of CPB larvae were assayed. Second and third instar larvae fed on transformed leaves molted earlier and had higher relative growth and consumption rates than larvae fed on nontransformed leaves, while efficiency of food utilization was unaffected. In contrast, fourth instar maximum weight gain and amount of leaves consumed were about 20% lower for the larvae fed on transgenic potato. Analysis of total protease activity of third instar larvae revealed reduction in overall proteolytic activity measured by azocasein hydrolysis, accompanied with inhibition of cysteine proteinase activity 24 h after ingestion of potato leaves expressing OCI and OCII. However, after long‐term feeding on transformed leaves proteolytic activities of larvae became similar to the controls. Although feeding on OCI/OCII leaves did not affect larval survival, coexpression of OC genes reduced the development time and thus significantly decreased plant damage caused by CPB larvae.     

Onshore–offshore variations of copepod community in northern South China Sea

Previous studies which have tested the feeding preferences of shredders for fungal species and the food quality of fungi used detritus uniformly colonized by a fungus, which is not the case for decaying leaves in streams. It is not known whether shredders in different development stages exhibit variations in feeding preference and larval performance. This study examined the feeding preferences and the growth of the third and the fifth instars of Pycnopsyche gentilis larvae using fungal-colonized patches and whole leaves, respectively, having different fungal species compositions (Alatospora acuminata, Anguillospora filiformis, Articulospora tetracladia, Tetrachaetum elegans, and all species combined). The aquatic hyphomycetes used were co-dominant on leaves in the stream inhabited by the caddisfly. During 14 d of feeding, the larvae of both instars did not show significant differences in feeding preferences for the patches growing on oak leaves, although the third instar larvae were slightly more selective than the fifth instar larvae. When fed with maple leaves for 18 d, larval growth rates, gross growth efficiencies, and survivorship were not significantly different among the fungal treatments. However, the larval growth of both instars fed with fungal-colonized leaves was always significantly greater than the growth of larvae fed with diets of uncolonized leaves. The third instar larvae grew faster than the fifth instar larvae, but the growth efficiencies of the two instars were similar. These results suggest that P. gentilis larvae exhibit less selectivity in their feeding than other caddisfly shredders that have been examined and that the dominant fungi colonizing leaves in their habitat are similar in palatability and food quality for this shredder. Handling editor: B. Oertli     

Antijuvenile influence of the precocene on the development of adult antennae in the large fruit-tree tortrix Archips podana Scop. (Lepidoptera: Tortricidae)

The influence of precocene II, an antijuvenile agent, on the development of adult antennae in the large fruit-tree tortrix A. podana Scop. was demonstrated. Treatment of the fifth instar larvae and prepupae with different doses of precocene proved to cause different sensitivity of the specimens to the juvenile hormone deficit. Treatment with 450 and 600 ??g precocene per specimen during the first days after ecdysis to the fifth instar caused the death of larvae. Treatment with 300, 450, and 600 ??g per specimen on the third day of the fifth instar larvae and prepupae caused a delay in the development of adult antennae. The results are discussed with respect to the role of the juvenile hormone in the development of imaginal structures during metamorphosis.     

A possible therapy by juvenile hormone I to Spodoptera littoralis (Boisd.) larvae previously treated by precocene II

An effective constant dose (55 μg) of precocene II (PII) was topically tested against the last three instars of Spodoptera littoralis (Boisd.) larvae. Application of PII induced morphogenetic abnormalities typical of juvenile hormone (JH) excess. The resultant imperfect insects included larval‐pupal mosaic and partial or severe cases of untanned pupae. The sixth‐instar larvae were more sensitive to PII administration than the two preceding instars. However, sensitivity of the last larval instar to PII varied with the timing of dose application relative to the developmental status of the larvae. Whereas the newly ecdysed (0‐day old) larvae were more sensitive, the older larvae of the same sixth‐instar showed sharp decrease in their sensitivity to PII with a concomitant increase of their age. Application of a single dose (5 μg) of JH I to PII pre‐treated larvae significantly (P < 0.001) reduced the production of imperfect insects where many PH‐treated larvae developed successfully to apparent normal pupae. Although a single dose of PII was more effective on S. littoralis larvae than repeated daily doses, the effectiveness of JHI‐therapy to PH pre‐treated larvae by repeated doses was less effective in producing perfect insects than JH‐therapy to PII pre‐treated larvae by single doses. The reversal of any of these by applied JHI is quite interesting but the mechanisms remain to be unraveled.     

Characterization of the Mamestra configurata (Lepidoptera: Noctuidae) larval midgut protease complement and adaptation to feeding on artificial diet,Brassica species,and protease inhibitor

The midgut protease profiles from 5th instar Mamestra configurata larvae fed various diets (standard artificial diet, low protein diet, low protein diet with soybean trypsin inhibitor [SBTI], or Brassica napus) were characterized by one‐dimensional enzymography in gelatin gels. The gut protease profile of larvae fed B. napus possessed protease activities of molecular masses of approximately 33 and 55 kDa, which were not present in the guts of larvae fed artificial diet. Similarly, larvae fed artificial diet had protease activities of molecular masses of approximately 21, 30, and 100 kDa that were absent in larvae fed B. napus. Protease profiles changed within 12 to 24 h after switching larvae from artificial diet to plant diet and vice versa. The gut protease profiles from larvae fed various other brassicaceous species and lines having different secondary metabolite profiles did not differ despite significant differences in larval growth rates on the different host plants. Genes encoding putative digestive proteolytic enzymes, including four carboxypeptidases, five aminopeptidases, and 48 serine proteases, were identified in cDNA libraries from 4th instar M. configurata midgut tissue. Many of the protease‐encoding genes were expressed at similar levels on all diets; however, three chymoptrypsin‐like genes (McSP23, McSP27, and McSP37) were expressed at much higher levels on standard artificial diet and diet containing SBTI as was the trypsin‐like gene McSP34. The expression of the trypsin‐like gene McSP50 was highest on B. napus. The adaptation of M. configurata digestive biochemistry to different diets is discussed in the context of the flexibility of polyphagous insects to changing diet sources. Published 2010 Wiley Periodicals, Inc.     

Preventive effects of N‐acetyl‐l‐cysteine against imidacloprid intoxication on Bombyx mori larvae

Imidacloprid, a widely used neonicotinoid insecticide, is toxic to silkworm (Bombyx mori). To explore whether N‐acetyl‐l ‐cysteine (NAC) has an effect on preventing silkworm (B. mori) from toxification caused by imidacloprid, we fed the fifth‐instar larvae with mulberry leaves dipped in 200 mg/L NAC solution before exposing in imidacloprid, and investigated the silkworm growth, survival rate, feed efficiency, cocoon quality, and the activities of antioxidant enzymes in midgut. The results showed that addition of NAC could significantly increase body weight, survival rate, and feed efficiency of imidacloprid poisoned silkworm larvae (P < 0.05), as well as cocoon mass, cocoon shell mass, and the ratio of cocoon shell (P < 0.05). Furthermore, it could significantly promote the activities of the antioxidant enzymes including superoxide dismutase, catalase, and glutathione peroxide in the midgut of fifth‐instar larvae under imidacloprid exposure at the late stage of treatment. In addition, it also could downregulate the malondialdehyde content. The results of our findings proved that the added NAC may have some beneficial effects on protection or restoration of antioxidant balance in imidacloprid exposed larvae.     

Comparative studies on pharmacokinetics and metabolism of the anti-juvenile hormone precocene II

The cuticular penetration and pharmacokinetics of the anti-juvenile hormone precocene II were determined in a sensitive species (Oncopeltus fasciatus) and an insensitive species (Heliothis zea). Precocene was sequestered by the fat body and slowly metabolized in Oncopeltus, but rapidly metabolized and excreted in Heliothis. Studies in vitro using inhibitors for cytochrome P-450 and for cyt P-450-NADPH-reductase, confirmed the anticipated detoxification of precocene by a mixed-function oxidase via the 3,4-epoxide. Use of the inhibitors in vivo had no influence on the metabolism of precocene.     

Treatment of larvae with repeated doses of precocene II induces precocious metamorphic changes in Spodoptera mauritia (Lepidoptera: Noctuidae)

In Spodoptera mauritia repeated daily treatments of larvae with 40 μg of precocene II throughout the fifth and sixth instar larval period had no effect on the larval-larval period but prolonged larval-pupal ecdysis. The resulting pupae showed precocious adult differentiation of mouth parts, wings, eyes, legs, and fat body.